Establishment Of An Indirect ELISA Antibody Detection Method For Porcine Pseudorabies Virus

As we know,pig pseudorabies virus disease is defined as a natural epidemic disease that is extremely difficult to prevent epidemics.It is widely spread on a global scale.Some countries in Europe and America already have effective disease prevention and control measures and culling procedures,but in third world countries,it is still difficult to effectively prevent and control the disease.In China,the pseudorabies epidemic situation was controlled by vaccine immunization.However,since 2010,the pseudorabies virus has undergone a wide range of mutations in China,As a result,the effect of prevention and control measures established in China before2010 has dropped significantly,causing the disease to re-emerge in the country.In order to meet the needs of new epidemic situation caused by high frequency mutation of pseudorabies virus?PRV?antigen in China,we designed and synthesized a number of peptides targeting the conserved antigenic epitopes of the glycoproteins gB,gC and gG of PRV with the help of bioinformatics analysis.The synthesized peptides,gB₈₇₂,gC₁₄₄,and gG₂₃₇,were used as coating antigens for this indirect ELISA.After optimization of coating antigen concentrations,blocking buffers,primary antibody dilutions,secondary antibody dilutions,and developing time,the indirect ELSIA kit for PRV antibody detections was assembled.The results showed that the ELISA was specific for detection of the antibodies against PRV,which had no cross-reaction with other five positive sera against swine fever virus,porcine reproductive and respiratory syndrome virus,foot-and-mouth disease virus?O type?,porcine parvovirus,porcine circovirus type 2.The comparative analysis revealed that the kit has specificity at 100%and sensitivity at 97%,suggesting that it can be used to detect the PRV antibody level in vaccinated or infected animals.The method was high duplicability with the less 10%variation of intra-and intro-assay coefficients,and the dilution of the PRV positive serum could reach 1:128.The indirect ELISA assay established in this study is of great significance for the investigations on vaccination evaluation,epidemiological analysis,and prevention and control of PRV.